Corporate Profile

Reinventing the Western Blot

ProteinSimple Retooled Blot Technology by Putting the Entire Workflow into a Capillary

One complete pathway in a single run: The NFκB cell signaling pathway was analyzed using a Sally Simple Western instrument. Each 5 µL sample of whole cell (WC) or nuclear extract (NE) lysate from either untreated (-) or TNFα-treated (+) cells was screened with eight different primary antibodies in replicate.

That conviction helped Harkness and his team build a company from 30 employees, no commercial products, and no revenue in 2008 to an organization that today has 170 employees, four product lines, and $50 million in revenue. “We are cash-flow positive and fully self-sustaining at this point. We fund all of our product development and growth capital needs with internally generated cash flow,” he says.

The company spent its first four years, from 2004 to 2008, developing the technology behind the Simple Western™ platform. When Harkness and his team joined the company, they initially focused on launching the first product, the NanoPro™ 1000, which separated proteins by charge, and targeted protein-signaling applications. They realized the technology platform could address a much larger market by separating proteins by size, a technique that is the basis of the Western blot. During the next few years, the team focused on using the Simple Western platform to reinvent and revolutionize the Western blot.

The company was able to reimagine the Western blot because Simple Western’s team approached Western blots from an entirely new angle. Instead of automating the movement of gels and blots typically used in a Western, the team put the entire workflow into a capillary.

“We were able to develop a fully automated, quantitative, reproducible immunoassay in a capillary for the first time in history,” Harkness says. “This allowed us to address all of the shortcomings associated with the 30-year-old Western blot technology.”

The Western blot is the most widely used protein analysis technique. “It was invented in the 1970s and has been done in much the same manual, tedious, multistep way ever since,” Harkness explains.

Traditionally, Western blot analysis has been a time-consuming, labor-intensive process with multiple steps that often required optimization and sometimes lengthy incubation times. The hands-on nature of the process makes it easy to introduce errors, and results often are considered difficult to reproduce. “Traditional Western blot analysis is semi-quantitative at best and is not easily reproducible from one day to the next or from one person to the next,” Harkness points out.

Although other companies had automated some of the more tedious steps of the process, none had developed a completely automated system—until ProteinSimple launched the Simple Western. By moving Western blot analysis from gels to capillaries and completely automating analysis, ProteinSimple says it is revolutionizing Western blot technology. Harkness describes the Simple Western as “a gel-free, blot-free, hands-free assay.”

For the first time, researchers can get quantitative, reproducible Western data automatically,” says Harkness. “Using Simple Western, the researcher prepares the sample as usual, pipettes it into a plate, places that plate in the instrument, presses ‘start,’ and walks away.” Fully analyzed, quantitative, reproducible data is ready within a few hours or, for large quantities of samples, overnight.

“This frees researchers’ time [and] produces higher quality data and more reproducible results.” With as little as 30 minutes of hands-on time, a researcher can now generate 96 Western data points and achieve coefficients of variation below 10%.

More than 35,000 researchers around the world are running Western blots today. “This is a billion dollar opportunity, and our goal is to target that entire market with a variety of options and price points,” Harkness says.

Biologics Are a Huge Opportunity

ProteinSimple also has developed a platform of tools focused on protein identification and characterization. “We have a set of products specifically for biologics,” Harkness says. These tools determine protein purity or detect protein aggregation in process development and in quality control. More than 800 systems have been sold globally. “Protein therapeutics developers use our tools because they work the same way every time [and] are robust and economical,” he adds.

For example, the iCE™ imaging capillary electrophoresis family of products is used for monitoring protein charge heterogeneity and, therefore, protein purity. This helps researchers ensure that the biologics produced are what they intended. That assurance speeds development and also provides sample analysis with high resolution and reproducibility.

A third analysis tool, MFI™, uses micro-flow imaging to characterize and quantify microscopic particles in formulations. It enables biologics developers to detect and visualize protein aggregates and particulates, as well as foreign contaminants like silicon oil droplets and air bubbles. The MFI 5000 series helps protein developers meet FDA requirements to quantify aggregates and particulates in biotherapeutic formulations. All of the company’s products are available with CFR Title 21 Part 11-compliant software, allowing assays to be validated to that standard.

ProteinSimple’s overaraching strategy is to be the “go-to” tool provider for proteins. With that in mind, it also offers a line of traditional gel and blot-imaging systems that provide high-sensitivity imagers for proteomic research from entry-level to high-throughput multiplexed systems.

First Is Never Easy

Harkness characterizes ProteinSimple as a relatively small company, but “a significant player in the markets in which we compete. We’ve gone after the areas in which we can make a difference.”

The challenge for ProteinSimple, and for any company with any breakthrough technology, is to break through the wall of resistance epitomized by the phrase, “We’ve always done it this way.” As Harkness acknowledges, “That wall is real, no matter what you sell. It’s hard for innovative companies to convince people to change, especially when the innovator is small and new. But, when we can talk with researchers and show them real results, they will make that change.” In the few years since the Simple Western has been available, more than 250 instruments have been installed in labs throughout the world.

Goal: Continued Growth

Before joining ProteinSimple, Harkness was CFO at Molecular Devices, where he helped company revenues grow from $38 million to $185 million in nine years. That company eventually sold for more than $600 million. So, when Harkness joined ProteinSimple in 2008, four years after its founding, he was looking for a company with good growth potential. In evaluating the young company, he recalls, “The technology underlying its products was important and broadly useful, and there was a lack of companies focused on protein analysis.”

Harkness says his management strategy is simply to grow the company. “Our goal is to build a $100 million protein analysis pure play. We’ve had good success and we want to continue it.”

ProteinSimple

Focus: ProteinSimple is focused on the development and manufacture of next-generation tools to measure and characterize proteins. Its goal is to deepen understanding of proteins and their roles in disease.

Jobs

GEN Jobs powered by HireLifeScience.com connects you directly to employers in pharma, biotech, and the life sciences. View 40 to 50 fresh job postings daily or search for employment opportunities including those in R&D, clinical research, QA/QC, biomanufacturing, and regulatory affairs.

If you have any questions about your subscription, click
hereto email us or call at (914) 740-2189.

You may also be interested in subscribing to the GEN magazine, an indispensable
resource for everyone involved in the business of translating discoveries at the
bench into solutions that fight disease and improve health, agriculture, and the
environment. Subscribe
today to see why over 60,000 biotech professionals read GEN to
keep current in the areas of genomics, proteomics, drug discovery, biomarker discovery,
bioprocessing, molecular diagnostics, collaborations, biotech business trends, and
more.